Generation of Tissue Engineered Nerve Grafts from GalSafe Porcine Neurons

项目总结 组织工程医疗产品有巨大的希望来解决目前无法修复的 神经创伤和/或神经退行性疾病。Axonova Medical的主导产品--组织工程神经 移植物(Teng)，是一种具有生物活性的“活支架”，可促进神经快速再生和功能恢复 当被植入修复主要周围神经损伤(PNI)时。以实现神经元活细胞为起点 生物量，Axonova常规的生物制造由活的，长的，排列的轴突束组成，穿过 轴突“拉伸生长”的专有过程。到目前为止，腾格被证明一直在提拔主持人 与目前的临床实践相比，轴突再生和改善功能恢复在大小两方面都是如此 PNI动物模型，包括修复5 cm节段缺损区。为了转移到制造业- 符合要求的生物量，Axonova已与Revivicor合作，利用它们的遗传基因产生GalSafe®TNG 改造猪神经元，以减轻最终在人类体内移植时的宿主免疫反应。 通过第一阶段的努力，GalSafe®Tengs成功地进行了拉伸种植，以生产出长度为 1厘米、3厘米和5厘米，健康和轴突密度达到或超过既定的质量基准。 此外，将1厘米的TENG移植到啮齿动物PNI模型中，以展示再生效果， 揭示GalSafe®Tengs存活并通过专有技术积极推动宿主轴突再生 TENG的作用机制，被称为轴突促进轴突再生(FAR)。与选择一起 作为一种安全和容易获得的起始生物质，其他产品开发挑战包括生物保存和 质量保证标准，以延长产品的保质期、运输和维持效力。 因此，当前的第二阶段SBIR计划解决了临床级生物制造的一个关键方面， 具体地说，验证有效的储存方案和确保产品健康、无菌、 和力量。TNG将从符合cGMP的细胞来源(GalSafe®猪神经元)中提取，冷- 使用符合cGMP的保存协议存储，然后在PNI的小动物模型和大动物模型中进行测试 来证明再生的功效。具体地说，Axonova将系统地完成这一目标 计划涉及三个具体目标：(1)确定新鲜GalSafe®tengs在已建立的猪身上的有效性 PNI模型；(2)有效的生物储备GalSafe®Tengs，以延长货架期并验证非侵入性释放 标准；以及(3)确定生物保存的GalSafe®Tengs在啮齿动物体内的效力和疗效 猪PNI模型。这个项目解决了下一代组织翻译中的一个主要挑战 通过验证活生物的制造、储存和释放标准来设计医疗产品 最终能够在人体上进行测试的产品。能够将GalSafe®Teng存储和分发为 货架“产品将通过降低成本和扩大患者接触范围极大地增加商业潜力，从而 改善患有严重PNI虚弱影响的患者的康复和生活质量。

PROJECT SUMMARY Tissue engineered medical products have tremendous promise to address the currently irreparable effects of neurotrauma and/or neurodegenerative diseases. Axonova Medical’s lead product, the tissue engineered nerve graft (TENG), is a bioactive “living scaffold” proven to promote rapid nerve regeneration and functional restoration when implanted to repair major peripheral nerve injury (PNI). Implementing living neuronal cells as the starting biomass, Axonova routinely biomanufactures TENGs comprised of living, long, aligned axon tracts through the proprietary process of axon “stretch-growth”. To date, TENGs have been shown to consistently promote host axon regeneration and improve functional recovery compared to current clinical practices in both small and large animal models of PNI, including the repair of 5 cm segmental defects. In order to move to a manufacturing- compliant biomass, Axonova has partnered with Revivicor to generate GalSafe® TENGs using their genetically engineered porcine neurons to mitigate a host immune response upon eventual transplantation in humans. Through the Phase I efforts, GalSafe® TENGs were successfully stretch-grown to create products at lengths of 1 cm, 3 cm, and 5 cm with a health and axon density that met or exceeded established quality benchmarks. Moreover, 1 cm TENGs were transplanted into a rodent PNI model to demonstrate regenerative efficacy, revealing that GalSafe® TENGs survived as well as actively drove host axonal regeneration via the proprietary mechanism-of-action of TENGs, referred to as axon facilitated axon regeneration (AFAR). Along with choosing a safe and readily available starting biomass, other product development challenges include biopreservation and quality assurance criteria to allow increased shelf life, transportation, and maintained efficacy of the product. Accordingly, the current Phase II SBIR program addresses a key facet of clinical grade biomanufacturing, specifically the validation of an effective storage protocol and release criteria ensuring product health, sterility, and potency. TENGs will be biofabricated from a cGMP-compliant cell source (GalSafe® porcine neurons), cold- stored using a cGMP-compliant preservation protocol, and then tested in small and large animal models of PNI to demonstrate regenerative efficacy. Specifically, Axonova will systematically complete the objectives of this program across 3 Specific Aims: (1) determine the efficacy of fresh GalSafe® TENGs in an established porcine PNI model; (2) effectively biopreserve GalSafe® TENGs to increase shelf-life and validate non-invasive release criteria; and (3) determine the potency and efficacy of biopreserved GalSafe® TENGs in vivo in rodent and porcine PNI models. This program addresses a major challenge in the translation of next-generation tissue engineered medical products by validating manufacturing, storage and release criteria for a living biological product to enable eventual testing in humans. The ability to store and distribute GalSafe® TENGs as an “off-the- shelf” product will vastly increase commercial potential by reducing costs and expanding patient access, thereby improving recovery and quality of life for patients suffering from the debilitating effects of major PNI.